Open AccessCharacterization of Polyethylene Glycol‐Polyethyleneimine as a Vector for Alpha‐Synuclein siRNA Delivery to PC12 Cells for Parkinson's Disease
Author(s) -
Liu YunYun,
Yang XingYi,
Li Zhong,
Liu ZhongLin,
Cheng Du,
Wang Yong,
Wen XiaoJun,
Hu JingYang,
Liu Jun,
Wang LiMin,
Wang HuiJun
Publication year - 2014
Publication title -
cns neuroscience and therapeutics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.403
H-Index - 69
eISSN - 1755-5949
pISSN - 1755-5930
DOI - 10.1111/cns.12176
Subject(s) - cytotoxicity , flow cytometry , transfection , mtt assay , viability assay , polyethylene glycol , gene delivery , microbiology and biotechnology , apoptosis , chemistry , peg ratio , zeta potential , intracellular , in vitro , biology , biochemistry , materials science , nanotechnology , gene , finance , economics , nanoparticle
Summary Aims Gene therapy targeting the SNCA gene yields promising results in the treatment of Parkinson's disease ( PD ). The most challenging issue of the RNA i gene therapy strategy is maintaining efficient delivery without inducing significant toxicity and other adverse effects. This study aimed to characterize polyethylene glycol‐polyethyleneimine as a vector for alpha‐synuclein si RNA delivery to PC12 cells for Parkinson's disease. Methods The characteristics of PEG‐PEI /si SNCA were analyzed via gel retardation assay and assessments of particle size and zeta potential. MTT cytotoxicity assay and flow cytometry were used to detect cytotoxicity and transfection efficiency in PC12 cells. Confocal laser scanning microscopy was employed to examine the intracellular distribution of PEG‐PEI / FITC ‐si SNCA after cellular uptake. RT‐PCR and western blotting were used to measure SNCA expression. The MTT cytotoxicity assay was used to study the effect of PEG‐PEI /si SNCA on cell viability. The protective effect of PEG‐PEI /si SNCA on MPP +‐induced apoptosis in PC12 cells was examined via flow cytometry and Hoechst staining. Results PEG‐PEI /si SNCA complexes were well‐developed; they exhibited appropriate particle sizes and zeta potentials at a mass ratio of 5:1. In vitro , PEG‐PEI /si SNCA was associated with low cytotoxicity and high transfection efficiency. Complexes were capable of successfully delivering si SNCA into PC12 cells and releasing it from the endosome. Furthermore, PEG‐PEI /si SNCA could effectively suppress SNCA m RNA expression and protected cells from death via apoptosis induced by MPP + . Conclusions Our results demonstrate that PEG‐PEI performs well as a vector for alpha‐synuclein si RNA delivery into PC12 cells. Additionally, PEG‐PEI /si SNCA complexes were suggested to be able to protect cells from death via apoptosis induced by MPP + . These findings suggest that PEG‐PEI /si SNCA nanoparticles exhibit remarkable potential as a gene delivery system for P arkinson's disease.